The Japanese journal of experimental medicine最新文献

Oxygen free radical (OFR) generation capacity of peritoneal macrophages was studied by chemiluminescent technique. Chemiluminescent (CL) response of macrophages from control, infected and immunized-infected mice was observed using non specific (Latex) and specific (S. typhi, cells and porins) stimulants at different time intervals. CL response was found to be significantly higher in immunized-infected group throughout the study period using all the three stimulants as compared to that in the infected as well as uninfected control mice. The mode of action of porin vaccine in increasing capacity of generating OFR is probably through increased expression of porin (protein) as well as carbohydrate receptors on the macrophage surface which leads to the stimulation of the whole caseade of respiratory burst or through the increase in the respiratory burst enzyme activities linked with each receptor or both.

We have generated a monoclonal antibody against prostatic adenocarcinoma, PC-Ab (IgM) was derived from a fusion using the fresh prostatic tissue of adenocarcinoma as the immunogen. Initial screening was performed by enzyme-linked immunosorbent assay (ELISA) on the soluble fraction of the immunogen. The specific analysis was performed by the avidin-biotin-complex immunoperoxidase method on paraffin-embedded sections of normal, benign and malignant neoplastic tissues from the prostate and various organs. PC-Ab reacted with well differentiated adenocarcinoma (83.3%), moderately differentiated adenocarcinoma (92.2%), and poorly differentiated adenocarcinoma (90.2%), respectively. According to classification by stage, the reaction rates with stage T0, T1, T2, T3 and T4 were 77.3%, 80.0%, 95.2%, 91.3% and 92.9%, respectively but no significant differences in the stages were seen among these groups. PC-Ab reacted with the epithelium of the normal prostate and benign prostatic adenoma, and human fetal tissues. Molecular weight and isoelectric point of the antigen recognized by this PC-Ab was estimated to be 57,000 daltons and 7.0, respectively. These results indicate that PC-Ab reacts with the antigen associated with human prostatic adenocarcinoma.

The specificity of five monoclonal antibodies, three raised against hepatitis B surface antigen (HBsAg) particles and two against envelope polypeptides, was tested for on a panel of 366 sera containing HBsAg of various subtypes (131 adw, 146 adr, 39 ayw and 50 ayr). Three monoclonals bound to HBsAg irrespective of subtypes, and therefore, were directed to the common antigenic determinants of HBsAg. Of these, two raised against particles (No. 824 and No. 7922) did not bind with reduced HBsAg particles. The other raised against peptides (No. 5124) bound to reduced HBsAg particles. It did not, however, bind to reduced and alkylated HBsAg particles, thereby indicating that it was directed to an epitope involving cysteine residues not contributing to the conformation. The remaining two monoclonals were directed to subtypic determinants not identical to any of d, y, w and r determinants. The subtypic determinant detectable by one of them (No. 4403), raised against HBsAg polypeptides, markedly increased after reduction of HBsAg particles with or without alkylation. In contrast, the subtypic determinant, detectable by the other monoclonal (No. 2155) raised against particles, substantially decreased after reduction. Non-identity of common or subtypic determinants detectable by the five monoclonals were established by blocking tests in which labeled antibody was competed by non-labeled antibody, of a homologous or heterologous specificity, for the binding with HBsAg. These monoclonals would be useful in studies for immunochemical configuration of HBsAg particles and epidemiology of novel subtypic determinants.

The renal brush border membrane vesicles (BBMV) were used to elucidate the early biochemical functional status during the course of experimental M. leprae infection in mice. The activities of the characteristic brush-border enzymes viz: alkaline phosphatase, leucine amino peptidase and gamma-glutamyl transpeptidase were found to be significantly decreased (p less than 0.001) at 3 and 6 months after infection. The transport of nutrients viz: D-glucose, L-alanine, L-lysine and L-aspartate across BBMV showed similar pattern. The activity of brush border enzymes and transport of nutrients across the membrane returned to normal at 9 months post-infection suggesting regeneration of the brush border membrane.

The morphological patterns of rejection of the heart allografts which were combinedly transplanted with skin grafts was examined to determine whether it was different from that of the heart grafts transplanted alone. The results showed that there were distinct differences in the pathology of the rejection between the two groups because in the single heart grafts mononuclear cells concentrated in almost all layers of myocardium but in the combinedly transplanted heart grafts not only concentration of them but also severe hemorrhage and edema were observed in it, and it suggests that skin graft has effects on rejection process of heart grafts. As we used an outbred strain of rats some survived heart grafts were observed and their pathology was characteristic in the interstitial accumulation of hypertrophic mast cells, most of which were degranulating.

To investigate genetic and sex factors in the local tumorigenesis by 7,12-dimethylbenz(alpha)anthracene (DMBA) in the mammary gland, both males and females of two different inbred strains of rats, Wistar/Furth (WF) and Copenhagen (COP), were subjected to dusting with approximately 1 mg of DMBA powder directly onto the exposed inguinal mammary tissue at 30 days of age. Locally growing tumors to 2 cm in mean diameter were harvested during 28 weeks after the carcinogen application. The incidence of macroscopic tumors of mammary origin was 100% in 15 WF females, 19% in 16 WF males, 85% in 20 COP females, and 74% in 19 COP males. Histologic pictures indicated the carcinomatous pattern composing mainly of differentiated adenocarcinoma of ductular cells in 12 tumors (80%) from WF females but not in any tumors from the other groups. On the other hand, they showed the sarcomatous pattern characterized by undifferentiated sarcoma of stromal cells in 2 tumors (13%) from WF females, 3 tumors (19%) from WF males, 16 tumors (80%) from COP females, and 14 tumors (74%) from COP males. The other 2 tumors from 1 WF and 1 COP females revealed the carcinosarcomatous pattern. Therefore, mammary ductular cells of WF are highly susceptible to DMBA and may be modified by sex factors in their carcinogenesis. Mammary stromal cells of COP are extremely susceptible to DMBA independently of sex factors.

The 5'-noncoding region of hepatitis C virus (HCV) genomes is highly conserved. A two-stage polymerase chain reaction (PCR), involving two pairs of primers deduced from the 5'-noncoding region of the HCV genome, was developed for a sensitive and specific detection of HCV RNA. The first stage of PCR was performed for 35 cycles with primers capable of multiplying fragments of 221 base pairs. PCR products in samples negative for HCV RNA were subjected to the second stage of PCR for 30 cycles with primers located internal to those employed in the first stage of PCR. The two-stage PCR detected up to 10 chimpanzee infectious doses/ml of HCV, and HCV RNA in 11 (92%) of 12 sera from patients with chronic non-A, non-B hepatitis without detectable antibodies to HCV by a commercial assay kit. Primers from the 5'-noncoding region of the HCV genome would be suitable for detecting HCV RNA by PCR, since the other regions of the HCV genome diverge extensively in sequence because of its nature as an RNA virus.

Filarial glomerulonephritis was studied using Dirofilaria immitis infected dogs. Of 34 infected dogs examined, 15 dogs (44.1%) had histopathological lesions in the kidney. These lesions included an increased number of mesangial cells and increased thickness of the matrix, the infiltration of the small round and plasma cells into the interstitium and thickening of the basement membrane. Deposits of IgG were demonstrated in the infected dogs, whereas C3 deposits were found in all dogs. Combined immunoglobulin and complement deposits were not always found in the dogs with histopathological lesions. The mean concentration (expressed as absorbance) of circulating immune complexes (CIC) was 0.675 +/- 0.517 in infected dogs, and 0.132 +/- 0.092 in uninfected dogs. Although there was significant difference in the level of CIC between infected and uninfected dogs (P less than 0.001), 11 dogs (32.4%) in infected group were negative. Otherwise, the CIC levels were correlated to the adult worm burden (r = 0.848; P less than 0.001) but not to the number of circulating microfilariae (mf) (r = 0.398; P less than 0.05). Transfer of mf to 7 naive dogs was performed to clarify the role of mf in the pathogenesis of filariasis. Antibodies to crude mf antigen became detectable two weeks after the transfer. Neither pathologic findings nor deposits of IgG and C3 in the kidney were found in dogs examined 20 days or 70 days after transfer. There was no evidence that histopathological lesions were induced by live mf, suggesting that adult worm burdens may be more closely related to filarial nephropathy.

To examine the roles of oxygen free radicals in the inhibition of tumor cell proliferation, we have used disulfiram (DS), a metal-chelator, to inactivate superoxide dismutase and ascorbic acid (AsA) to inhibit catalase. Simultaneous addition of DS and AsA to Meth A tumor cells or Ehrlich ascites tumor cells induced marked inhibition of the cell proliferation assessed by [3H]thymidine uptake and trypan blue dye exclusion method. Similar augmented inhibition of Meth A tumor cell proliferation was observed when the cells were pretreated with DS and AsA. However, the addition of catalase (2000 u/ml) nullified the augmentation of anti-proliferative effect which can be induced in the combined use of DS and AsA. These results suggest that the steady-state increase of intracellular oxygen free radicals within tumor cells could be induced in the combined use of DS and AsA.

IgM and IgG natural antibodies to Tamm-Horsfall glycoprotein (THP) were found in serum samples of all healthy individuals tested by the ELISA technique. The IgM anti-THP antibody level was higher in the group 1-20 years old than the IgG anti-THP. The IgG anti-THP rose with increase in age (greater than 21 years old groups) and then the IgG and IgM anti-THP activity over aging remained constant. The natural anti-THP antibodies possess a lower degree of specificity and/or avidity than induced antibodies. The antibody titers against THP determined in 61 adult patients with chronic kidney diseases was significantly lower than that in adult controls. This low level of naturally occurring THP antibodies appears to be a general phenomenon. In these patients, diminished antibody levels appeared against a panel of self (collagen, fibronectin, THP) and non self (bovine gamma globulin (BGG), ovalbumin (OVA)) antigens as compared with normal controls. The low levels of these antibodies are not associated with a concomitant drop of IgG and IgM in their sera.